中国水稻科学 ›› 2023, Vol. 37 ›› Issue (4): 359-367.DOI: 10.16819/j.1001-7216.2023.221113

• 研究报告 • 上一篇    下一篇

定向敲除SD1基因提高水稻的抗倒性和稻瘟病抗性

李刚1, 高清松2, 李伟2, 张雯霞2, 王健1, 程保山1, 王迪1, 高浩1, 徐卫军1, 陈红旗3(), 纪剑辉2()   

  1. 1江苏徐淮地区淮阴农业科学研究所/淮安市农业生物技术重点实验室,江苏 淮安 223301
    2淮阴师范学院/江苏省环洪泽湖生态农业与生物技术重点实验室, 江苏 淮安 223300
    3中国水稻研究所 水稻生物育种全国重点实验室, 杭州 311401
  • 收稿日期:2022-11-24 修回日期:2023-02-08 出版日期:2023-07-10 发布日期:2023-07-17
  • 通讯作者: *email: chqhzfy@126.com;jijianhui@hytc.edu.cn
  • 基金资助:
    淮安市自然科学基金项目(HAB202076);江苏省重点研发计划资助项目(BE2021334);江苏省重点研发计划资助项目(BE2021323);国家自然科学基金资助项目(32070345);江苏省自然科学优秀青年基金项目(BK20180107)

Directed Knockout of SD1 Gene Improves Lodging Resistance and Blast Resistance of Rice

LI Gang1, GAO Qingsong2, LI Wei2, ZHANG Wenxia2, WANG Jian1, CHEN Baoshan1, WANG Di1, GAO Hao1, XU Weijun1, CHEN Hongqi3(), JI Jianhui2()   

  1. 1Huaiyin Institute of Agricultural Science in Xuhuai Region of Jiangsu/Huai'an Key Laboratory of Agricultural Biotechnology, Huai'an 223001, China
    2Jiangsu Key Laboratory for Eco-Agricultural Biotechnology Around Hongze Lake, Huaiyin Normal University, Huai’an 223300, China
    3State Key Laboratory of Rice Biological Breeding Biology, China National Rice Research Institute, Hangzhou 311401, China
  • Received:2022-11-24 Revised:2023-02-08 Online:2023-07-10 Published:2023-07-17
  • Contact: *email: chqhzfy@126.com;jijianhui@hytc.edu.cn

摘要:

【目的】 为改良高产粳稻品种淮119株高偏高及易感稻瘟病等不利性状,利用CRISPR/Cas9基因编辑技术对淮119中SD1基因进行定向敲除,为淮119后代品种改良奠定基础。【方法】 利用CRISPR/Cas9系统,以SD1基因为靶基因,构建基因敲除载体,以农杆菌介导转化淮119,获得无转基因插入的纯合突变株,进一步对纯合突变株株高、农艺性状、稻瘟病抗性及氮素吸收利用能力等进行综合分析。【结果】 利用农杆菌转化淮119,鉴定获得1株无转基因载体序列插入的纯合突变株。大田种植发现,野生型淮119出现60%面积以上的倒伏,而sd1纯合突变株群体由于株高变矮,从而有效避免了生育后期的倒伏。此外,用不同浓度的GA(0.01~1.00 μmol/L)处理,野生型苗高均显著高于突变株,表明sd1突变导致对外源GA敏感性降低。稻瘟病抗性鉴定结果表明,sd1基因突变不仅有效降低株高,同时增强了对稻瘟病的抗性水平。不利因素是,SD1基因的敲除降低了淮119的氮素吸收利用效率。【结论】 淮119中SD1基因定向敲除获得的无转基因插入纯合突变株,不仅株高显著下降,且稻瘟病抗性得到增强。

关键词: CRISPR/Cas9, 基因编辑, SD1, 稻瘟病, 抗倒性, 水稻

Abstract:

【Objective】 Huai 119 is a high-yielding japonica rice variety. To improve its unfavorable traits, especially high plant height and poor resistance to rice blast, we used the CRISPR/Cas9 gene editing technology to knock out its ‘Green Revolution’ gene SD1.【Method】 We selected the SD1 gene as the target to construct the CRISPR/Cas9 gene knockout vector, which was then transformed into Huai 119 by Agrobacterium-mediated transformation method, and the homozygous sd1 knockout line without transgenic insertion was obtained. Subsequently, we compared and analyzed the plant height, rice blast resistance, and nitrogen uptake and utilization efficiency of the sd1 and wild-type lines.【Result】 We successfully isolated a homozygous sd1 knockout line without transgenic insertion in the background of Huai 119. In field paddy, it was found that over 60% of the planting area of wild-type Huai 119 was lodging, while the sd1 homozygous mutant population effectively avoided lodging in the later stages of growth due to its shorter plant height. In addition, after treatment with different concentrations of GA (0.01-1.00 μmol/L), the plant height increase of wild-type plants was significantly greater than that of the sd1 line, indicating that the sensitivity of sd1 line to exogenous GA treatment was reduced. The identification of rice blast resistance shows that knocking out SD1 also contributed to the improvement of rice blast resistance of Huai 119. However, the absorption and utilization efficiency of nitrogen decreased due to the knockout of SD1. 【Conclusion】 The knockout of SD1 gene in the rice variety Huai 119 not only improves lodging resistance by reducing plant height, but also enhances the resistance to rice blast.

Key words: CRISPR/Cas9, gene editing, SD1, rice blast, lodging resistance, rice